Differentially Expressed Gene Screening Of Urine And Kidney Tissue Samples From Rats’ Renal Fibrosis Model And Bioinformatics Analysis

Renal fibrosis as the final outcome of many renal diseases,has always been paid more attention to by the researchers.To better understand how transcripts be a player in this process and to find biomarkers for renal fibrosis diagnosis,we compared transcriptome sequencing data on renal tissues and urine samples respectively between unilateral ureteral obstruction(UUO)and shamed(Sham)rat model.A serie experiments and analysises in or between groups confirmed that the experimental model had been constructed successfully and the sequencing data was reliable.Numerous genes including long noncoding RNAs(lncRNAs)with significant changes in their expression were identified through differential expression analysis.10 lnCRNAS were up regulated and 8 lncRNAs were down regulated in urines of the UUO rats.24 lncRNAs were up regulated and 79 lncRNAs were down regulated in the renal tissues of the UUO rats.The lncRNAs,which were expressed in statistical significance in the renal tissue,intersected with the IncRNA in the urine sample.5 lncRNA expression were up regulated and 2 down regulated.Several differentially expressed lncRNAs above-mentioned were confiremed by reverse transcription polymerase chain reaction(RT-qPCR).The verification results consisted with the sequencing results.Pre-miRNA prediction was conducted in the up regulated urine sample IncRNAs.LncRNA NONRATT042796 was the pre-sequence of rno-mir-207 which had the relationship with the TGF-β signaling pathway.Superfamily analysis confirmed that a several of lncRNAs are included in the TUG1 superfamily which promoted the occurrence of fibrosis.GO and KEGG analysis were conducted in the differential expression mRNAs in urine sample and renal tissue.A great quantity of them were associated with fibrosis growth factors stimulation,inflammatory reaction and autophagy-lysosome pathway.Up regulated lncRNA in renal tissues were conducted by cis-acting element analysis.Many target genes had the function of the occurrence and development of fibrosis.19 lncRNAs which upregulated in renal tissue of UUO rats were predicted containing several conserved Smad3 binding motifs in the promoter.Among them,lncRNAs with putative promoter containing more than 3 conserved Smad3 binding motifs were demonstrated to be induced by TGF-β significantly in NRK-52E cells.We further confirmed that lncRNA TCONS 00088786 and TCONS 01496394 were regulated by TGF-β stimulation and also could influence the expression of some fibrosis-related genes through a feedback loop.Based on the transcriptome sequencing data,RT-qPCR detection,cytology relevant experiments and bioinformatics analysis,we demonstrate that new biological effected transcripts in urine sample and renal tissue are picked up.These data provide new information about the involvement of transcripts,expecially lncRNAs,in renal fibrosis,indicating that they may serve as candidate biomarkers or therapeutic targets in the future.